Abstract
Osteosarcoma is a disease prone to recurrence and metastasis, and adenovirus expression vector is frequently studied as a therapeutic target of osteosarcoma in recent years. The present study attempts to explore the effect of adenovirus-mediated siRNA targetting ezrin on the proliferation, migration, invasion, and apoptosis of human osteosarcoma MG-63 cells. Human osteosarcoma MG-63 cell line was selected for construction of recombinant adenovirus vector. The mRNA and protein levels of ezrin, Bcl2-associated X protein (Bax), B cell lymphoma-2 (Bcl-2), p21, p53, Caspase-3, matrix metalloproteinase (MMP) 2 (MMP-2) and MMP-9, Cyclin D1, and cyclin-dependent kinase 4a (CDK4a) were determined. Through ELISA, the levels of Caspase-3, MMP-2 and MMP-9 were examined. Finally, human osteosarcoma MG-63 cell viability, growth, invasion, migration, and apoptosis were detected. Initially, adenovirus expression vector of ezrin was constructed by ezrin 2 siRNA sequence. Adenovirus-mediated siRNA targetting ezrin reduced expression of ezrin in MG-63 cells. The results revealed that adenovirus-mediated siRNA targetting ezrin elevated expression levels of Bax, p21, p53, and Caspase-3, Cyclin D1, and CDK4a and reduced expression levels of Bcl-2, MMP-2 and MMP-9. Furthermore, adenovirus-mediated siRNA targetting ezrin inhibited human osteosarcoma MG-63 cell viability, growth, invasion, and migration, and promoted apoptosis. Our study demonstrates that adenovirus-mediated siRNA targetting ezrin can induce apoptosis and inhibit the proliferation, migration, and invasion of human osteosarcoma MG-63 cells.
Highlights
Osteosarcoma is a kind of cancerous tumor in the bone, which is the commonest primary bone cancer [1,2]
The results showed that compared with ezrin 1 and ezrin 4, down-regulation of ezrin 2 and ezrin 3 siRNA was significant (P
The results of Western blot analysis were consistent with that of reverse-transcription quantitative PCR (RT-qPCR), which indicated that the recombinant adenovirus vector could mediate the mRNA and protein levels of ezrin in MG-63 cells (Figure 3)
Summary
Osteosarcoma is a kind of cancerous tumor in the bone, which is the commonest primary bone cancer [1,2]. Osteosarcoma occurs most widely amongst children and adolescents [3]. Osteosarcoma is hardly ever diagnosed before the age of 5 while its incidence increases significantly by the time of puberty, with its incidence peak during the teen years and the second peak occurs in the elderly (aged ≥60 years) [4]. Its occurrence is mainly influenced by sex, age, ethnicity, tumor size, genetic factors, and environmental insults [5]. It frequently occurs in other cancer patients after therapeutic radiation, in patients with some kind of cancer predisposition syndromes, as well as in individuals with Paget disease of the bone [6]. New treatment strategies for osteosarcoma are in urgent need [10]
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